Figure 4. LDLRAD2 activates Wnt/β-catenin signaling pathway in GC. Gene-set enrichment analysis (GSEA) indicated that LDLRAD2 expression was closely associated with Wnt/β-catenin pathway (A). Western blotting analysis showed that overexpression of LDLRAD2 significantly increased the nuclear expression of β-catenin, while silence of LDLRAD2 dramatically decreased the nuclear expression of β-catenin in GC cell lines (B). Top/Fop flash assay also demonstrated that overexpression of LDLRAD2 significantly activated Wnt/β-catenin signaling pathway, while silence of LDLRAD2 dramatically restrained Wnt/β-catenin signaling pathway (C). Immunofluorescence assay also showed that LDLRAD2 could promote the nuclear distribution of β-catenin in GC cells (D). Overexpression of LDLRAD2 significantly promoted the expression of downstream genes of Wnt/β-catenin signaling including c-myc, VEGF, Twist, and MMP7, while silence of LDLRAD2 inhibited the expression of these genes (E).